Effects of a bioartificial liver support system on acetaminophen induced acute liver failure canines

AIM To evaluate the safety and efficacy of the bioartificial liver support system in canines with acute liver failure (ALF). METHODS Nine canines with acute liver failure by acetaminophen induced received TECA Ⅰ bioartificial liver support system (BALSS) from Hong Kong TECA LTD Co. Blood was perfused through a hollow fiber tube containing (1 2)×10 10 the porcine hepatocytes. In contrast, another 10 canines with acute liver failure by Acetaminophen received drugs. Each treatment lasted 6 hours. RESULTS BALSS treatment resulted in beneficial effects for acetaminophen induced ALF canines with survival and with the recovery of the liver functions and tissues, and plasma ammonia decreased from 135 9μmol/L ± 17 5μmol/L to 65 7μmol/L ± 22 0μmol/L , 32 5μmol/L ± 8 8μmol/L , GPT from 97 8U/L ± 8 7U/L to 64 8U/L ± 11 9U/L , 19 0U/L ± 6 3U/L , GOT from 103 0U/L ± 16 7U/L to 75 7U/L ± 19 6U/L , 26 5U/L ± 5 0U/L , and AKP from 158 3U/L ± 12 1U/L to 114 5U/L ± 19 8U/L , 43 8U/L ± 5 6U/L during and after the treatment. In contrast, 10 ALF canines in both the drug and control groups died 1 or 2 days after treatment. CONCLUSION TECA 1 artificial liver support system is safe and efficacious for canines with acute liver failure.

Three-dimensional Localization of Impacted Canines and Root Resorption Assessment Using Cone Beam Computed Tomography

Summary： The purpose of this study was to develop a new way to localize the impacted canines from three dimensions and to investigate the root resorption of the adjacent teeth by using cone beam computed tomography （CBCT）. Forty-six patients undergoing orthodontic treatments and having impacted canines in Tongii Hospital were examined. The images of CBCT scans were obtained from KaVo 3D exam vision. Angular and linear measurements of the cusp tip and root apex according to the three planes （mid-sagittal, occlusal and frontal） have been taken using the cephalometric tool of the InVivo Dental Anatomage Version 5.1.10. The measurements of the angular and linear coordinates of the maxillary and mandibular canines were obtained. Using this technique the operators could envision the location of the impacted canine according to the three clinical planes. Adjacent teeth root resorption.of 28.26 % was in the upper lateral incisors while 17.39% in upper central incisors, but no lower root resorption was found in our samples. Accurate and reliable localization of the impacted canines could be obtained from the novel analysis system, which offers a better surgical and orthodontic treatment for the patients with impacted canines.

The Prevalence of Impacted Maxillary Canines in a Palestinian Population: A Retrospective Study

Objective: This study aims to estimate the prevalence of impacted maxillary canines using panoramic radiographs. Methods: A sample of 1321 panoramic radiographs were retrospectively examined for the presence of impacted maxillary canines. The radiographs were obtained from the records of a private dental clinic from the year 2009 till 2016. Results: The prevalence of impacted canines among the examined population (n = 1321) was found to be 1.8%. There was a significant association between gender and canine impaction (P 0.05) difference in the prevalence of right or left canine impaction was found. Conclusion: The prevalence of impacted maxillary canines (1.8%) in this study was within the range reported among other populations.

Short-term results in canines of novel stent-graft design for chimney technique in TEVAR

Introduction:Parallel stent-stent grafting is a major endovascular technique used to preserve the supra-aortic branches during thoracic endovascular aortic repair(TEVAR)of aortic pathologies involving the aortic arch.The short-and mid-term results of this technique are satisfactory;however,endoleak remains a major concern.Thus,here we designed a new chimney stent-graft to decrease the endoleak rate.Aim:To testify the feasibility and safety of the new chimney stent-graft system in a canine model.Material and methods:Six Labrador retrievers were used.Pre-operative data were collected and all operations were performed under general anesthesia.The main and chimney stent-grafts were implanted through the abdominal aorta and left subclavian artery approaches,respectively.Completion digital subtraction angiography(DSA)was performed to confirm the immediate outcomes.All dogs were fed separately for 6 months and sacrificed after aortic angiography.The thoracic aorta and the main and chimney stent-grafts were harvested for histopathologic examination.Results:No complications were found in follow-up DSA.All branch arteries were patent.In?ammatory responses were observed around the stent-grafts in 3 experimental animals,and slight hyperplasia was observed in the surrounding tissues compared with the normal vessels.There was no mural thrombus in the stent,endothelial cells were noted on the inner surface of the stent,and thrombus was formed in the outer skirt and gutter area.The histopathologic examinations revealed similar results to those of gross necropsy observations.Conclusions:This study demonstrated the feasibility and safety of the Longuette stent-graft and the first to report a revised stent-graft specific for chimney technique.

Changes in the crown angulation and dental arch widths after nonextraction orthodontic treatment: Model analysis of mild crowding with high canines

This study was undertaken to examine which factors contributed to the correction of crowding in two patients who underwent nonextraction orthodontic treatment. A study model analysis was conducted to determine the effects of the orthodontic treatment for crowding with high canines on crown angulation and dental arch width in two patients. The results showed that the crown angulation was significantly increased, indicating distal tipping in the maxillary dental arch. This tendency was most commonly observed in the premolars among the lateral teeth. With respect to the dental arch width, the largest change was evident in the first molar and first premolar regions in cases 1 and 2, respectively. On the basis of these results, up-righting of mesially tipped lateral teeth and expansion of narrow dental arches could prove to be the keys to the success of space regaining or correction of high canines and mild crowding.

Modified orthodontic treatment of substitution of canines by first premolars:A case report

BACKGROUND Canines are the most important teeth in the dentition.Usually,doctors choose to remove premolars rather than canines.Canine extraction is extremely rare in orthodontic treatment.However,dentists sometimes encounter situations in which canines require extraction due to defects caused by improper medical treatment.CASE SUMMARY The present study reports a case of a class II adult patient treated with the extraction of maxillary canines and right mandibular second premolar.After postactive treatment for 28 mo,then the canines were substituted by the upper first premolar,a satisfactory occlusal was established,the lips were competent,and the profile was improved.Intraoral pictures and X-ray data retrieved 3 years after the end of orthodontic treatment demonstrated the possibility of canine extraction and premolar substitution of canines in function and beauty.CONCLUSION The extraction of canines and substitution by first premolars could be a feasible orthodontic treatment.

Cone-Beam Computed Tomography Evaluation of the Positions of Unerupted Maxillary Canines with Incisor Root Resorption

Purpose: To establish a new coordinate system using the incisive canal and incisive foramen in cases confirmed to have root resorption in the maxillary incisor region by cone beam computed tomography (CBCT) to investigate the positions of the central and lateral incisor roots and erupting maxillary canine tooth crowns in the horizontal plane. Methods: Nine patients (two males;mean age: 10.5 years old) with suspected incisor root resorption due to erupting maxillary canines on panoramic X-ray images and in whom incisor root resorption was confirmed on CBCT images were evaluated. A control group of 12 patients with a supernumerary tooth on one side (three males;mean age: 8.6 years old) was also examined. X, Y, and Z-axes were defined, and the positions of the centers of the central incisor root (U1) and lateral incisor root (U2) and the canine cusp (U3) were examined, along with alveolar process width and length. Results: In the control group, U1, U2, and U3 were located within a certain range without overlap, while, in the incisor root resorption group, U3 overlapped with U1 and U2 and tended to deviate centrally. U2 tended to be located further posteriorly than U3. The anteroposterior diameter of the alveolar process was 1.2 mm shorter in the incisor root resorption group (p < 0.05). Conclusions: The risk of incisor root resorption accompanying canine eruption can be evaluated early by investigating the canine position on a horizontal plane established on the upper anterior tooth dentition CT images with a coordinate system using the incisive canal and incisive foramen.

Benefits of Using CBCT in Bilateral Maxillary Impacted Canines Extraction:A Case Report

Aim: Canines are the second most impacted teeth in jaws after third molars. Perforation of the anatomic structures, such as maxillary sinus and nasal fossa, are some of the complications of the surgery. In this case, bilateral maxillary impacted canines’ extraction, with the guidance of CBCT is done without any complication. Case: A 24-year-old female patient was referred to Istanbul University, Oral and Maxillofacial Surgery. Left and right deciduous canines and swelling at the right maxillary canine’s labial area were seen during the intraoral examination. Bilateral impacted canines were determined after evaluating the present CBCT images of the patient and orthodontic treatment was advised to the patient. Due to the refusal of orthodontic treament, extractions of the impacted teeth were decided. After the evaluation of CBCT images, the maxillary right canine was determined at the labial site while the maxillary left canine was determined at the palatinale site. Results: Right maxillary canine was lying between the right first incisor and right first molar and It was extracted with mucoperiosteal vestibule flap procedure. For the extraction, the tooth was divided into two pieces from the enamel-cement junction. Left maxillary canine was lying between the left first incisor and left first molar and It was extracted with mucoperiosteal palatinale flap procedure. Flap was stitched primarly with 3/0 silk sutures. Palatinal plaque, which was prepared before the surgery, was applied. Antibiotic, analgesic and moutwash were prescribed to the patient. 1 week following the surgery, stitches were removed and the healing was optimal. Conclusion: Impacted canines are a common clinic condition which requires a multi-discipliner approach. CBCT can increase the success of the treatment by preventing possible complications with determining the proximity of the anatomical structures and the localization of the impacted teeth. There were no sign of complication and the healing was optimal.

Porphyromonas gulae infection in canines,pet owners and veterinarians in China:an epidemiological study and risk factor analysis

Porphyromonas gulae is a clinically prevalent,anaerobic,oral bacteria in canines,that may be a causative agent of canine periodontal disease,and a potential threat to human oral health.Research on P.gulae pathogenicity in canines,their owners,and veterinarians is lacking in China.This study aimed to determine the isolation and detec-tion rates of P.gulae in gingival crevicular fluid(GCF)samples from 101 canines in Beijing,using anaerobic culture techniques and 16S rRNA gene sequencing.The main risk factors for the transmission of P.gulae from canines to humans were also analyzed through analyzing the statistical data on risk factor variables from 103 canine owners and 60 veterinarians in Beijing who tested positive for P.gulae detection in GCF samples.The isolation and detection rates of P.gulae in canines were 31.5%(29/92)and 92.1%(93/101),respectively,compared with detection rates of 24.3%(25/103)in canine owners,43.3%(26/60)in veterinarians,and 52.0%(13/25)in dentists.The degree of contact with canines(P=0.001,P<0.01)and smoking(P=0.021,P<0.05)were significant risk factors for P.gulae detection in owners.Moreover,the degree of contact during ultrasonic scaling(P=0.065,0.05<P<0.1)was the most important risk factor for the positive detection of P.gulae in veterinarians.These findings suggest that P.gulae may colonize the human oral cavity through intimate contact with canines or participation in dental ultrasonic scaling operations.

Three-dimensional-printed collagen/chitosan/secretome derived from HUCMSCs scaffolds for efficient neural network reconstruction in canines with traumatic brain injury

The secretome secreted by stem cells and bioactive material has emerged as a promising therapeutic choice for traumatic brain injury(TBI).We aimed to determine the effect of 3D-printed collagen/chitosan/secretome derived from human umbilical cord blood mesenchymal stem cells scaffolds(3D-CC-ST)on the injured tissue regeneration process.3D-CC-ST was performed using 3D printing technology at a low temperature(20C),and the physical properties and degeneration rate were measured.The utilization of low temperature contributed to a higher cytocompatibility of fabricating porous 3D architectures that provide a homogeneous distribution of cells.Immediately after the establishment of the canine TBI model,3D-CC-ST and 3D-CC(3D-printed collagen/chitosan scaffolds)were implanted into the cavity of TBI.Following implantation of scaffolds,neurological examination and motor evoked potential detection were performed to analyze locomotor function recovery.Histological and immunofluorescence staining were performed to evaluate neuro-regeneration.The group treated with 3D-CC-ST had good performance of behavior functions.Implanting 3D-CC-ST significantly reduced the cavity area,facilitated the regeneration of nerve fibers and vessel reconstruction,and promoted endogenous neuronal differentiation and synapse formation after TBI.The implantation of 3D-CC-ST also markedly reduced cell apoptosis and regulated the level of systemic inflammatory factors after TBI.

Establishment and characterization of a canine chondrosarcoma cell line:Mango

In the global progress of bone tumor research,established stable and long-lasting transgenic chondrosarcoma(CSA)cell lines are rare,mainly of murine and human origin,while the establishment of canine CSA cell lines has yet to be reported.This study established a canine CSA cell line to facilitate the basic clinical study of canine CSA.Fifty fve cases of canine osteolytic disease were collected,and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture.A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully cultured.According to the clinical characteristics of the dog and the histopathological results of the primary tumor,CSA was diagnosed,and the CSA cell line was designated Mango.Immunohistochemical(IHC)results showed that the immunoreactivity of bone gamma-carboxyglutamate protein(BGLAP),secreted protein acidic and rich in cysteine(SPARC),alkaline phosphatase(ALPL),vimentin(VIM)and S100 were positive.However,the immunoreactivity of pan-cytokeratin(PCK),chromogranin A(CGA),and platelet endothelial cell adhesion molecule-1(CD31)was negative.Immunofuorescence(IF)results showed that the protein expressions in the Mango cell line were consistent with the IHC identifcation of the primary tumor.The Mango cell line’s doubling time was 43.92 h,and the cell formation rate exceeded 20%.There were abnormal chromosome numbers,hetero staining with toluidine blue,and certain calcifcation abilities.It could be passaged stably and continuously without changing the cell morphology and characteristics.In vivo,the cells were successfully injected into the nude mice model with a tumorigenic rate of 100%.The immunophenotype of the xenograft tumor was consistent with that of the primary tumor.Therefore,we efectively established a canine CSA cell line.As a promising cell material,this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic research of canine CSA.Moreover,because of its similarity to human CSA,the animal model of CSA is also indispensable for investigating human CSA.

Molecular characterization of canine parvovirus type 2(CPV2)reveals a high prevalence of the CPV2c genotype among dogs sufering from diarrhea

Canine parvovirus 2(CPV-2)is a highly contagious virus in dogs that typically causes hemorrhagic enteritis and a high mortality rate in unvaccinated puppies.The genetic variability and antigenic diversity of CPV-2 hinder its efective prevention of infection by vaccination.To investigate the epidemiology and genetic characteristics of CPV-2 in China,rectal swabs from afected dogs were collected from diferent animal clinics in Kunshan from 2022 to 2023.Preliminary detection and capsid gene sequencing of CPV-2 were performed using previously described primers and protocols.The overall detection rate for CPV-2 was 16.5%(33/200).A signifcant association was found between the CPV-2-positivity and clinical signs,age,breed and vaccination status.Sequence analysis revealed the presence of CPV-2c genotypes in all positive samples,which were genetically similar to other Asian CPV-2c strains.Notably,four key mutations(A5G,F267Y,Y324I and Q370R)were detected in all isolates,and one novel mutation(I447M)was detected in three CPV-2 isolates.These mutations in the CPV-2 strains could impact vaccine efcacy and the efectiveness of the virus immune evasion.Surprisingly,no recombination events were observed between the identifed CPV-2c strains and reference strains from China.Our data revealed that amino acid residues 324,426 and 440 of VP2 may under strong selection pressure.This pattern of genetic variation in the CPV-2 lineage warrants continuous laboratory-based surveillance programs in other parts of China to better understand the pattern of seasonal distribution and association between emerging genotypes and the intensity of disease severity.

Preparation of anti-canine interleukin-31 receptor alpha polyclonal antibody and evaluation of its therapeutic efect in canine atopic dermatitis

Canine atopic dermatitis(CAD)is a prevalent genetically susceptible infammatory and pruritic allergic skin condition afecting not only the health of dogs but also the quality of life of their owners.Interleukin-31(IL-31)and interleukin-31 receptor alpha(IL-31RA)are essential for the development of pruritus in primates and mice.Hence,it is expected that inhibiting IL-31RA will be an efective approach to alleviate pruritus.The purpose of the study was to produce anti-canine IL-31RA polyclonal antibodies(anti-IL-31RA pAbs)and evaluate their efcacy in inhibiting house dust mite(HDM)-evoked pruritic responses.Dogs were immunized with antigens formed by IL-31RA recombinant short peptides coupled to BSA to produce anti-IL-31RA pAbs.The CAD model was developed by using HDM allergen stimulation,and the efects of IL-31RA pAbs on the reduction of pruritus in CAD model dogs were examined.The Canine Atopic Dermatitis Extent and Severity Index(CADESI)-4 and pruritus Visual Analog Scale(pVAS)were utilized to evaluate pruritic responses,and skin tissue samples were collected from the inguinal area for pathological assessment of skin infammatory cell infltration.The results showed that anti-IL-31RA pAbs with high titers(1:128,000)and specifcity were efectively produced.In the CAD model group,the severity of skin damage,pruritus score,infammatory cell infltration and level of infammatory factors were considerably elevated.Anti-IL-31RA pAbs relieved pruritic behavior and dermatitis in dogs compared to placebo-treated dogs.In conclusion,anti-IL-31RA pAbs efectively suppressed CAD in vivo and are anticipated to be an efective novel treatment for pruritic skin disorders such as CAD.

Emergence of the stromal vascular fraction and secretome in regenerative medicine

Recently,we read a mini-review published by Jeyaraman et al.The article explored the optimal methods for isolating mesenchymal stromal cells from adipose tissue-derived stromal vascular fraction(SVF).Key factors include tissue source,processing techniques,cell viability assessment,and the advantages/disadvantages of autologous vs allogeneic use.The authors emphasized the need for standardized protocols for SVF isolation,ethical and regulatory standards for cell-based therapy,and safety to advance mesenchymal stromal cell-based therapies in human patients.This manuscript shares our perspective on SVF isolation in canines.We discussed future directions to potentiate effective regenerative medicine therapeutics in human and veterinary medicine.

Clinicopathological Description of a Chronic Lymphocytic Leukemia in a Dog

Hematopoietic tumors are associated with diagnostic and therapeutic challenges and a poor prognosis. Furthermore, their pathophysiology in dogs is not well understood. Chronic lymphocytic leukemia has been rarely reported in golden retrievers, perhaps due to difficulties in diagnosis and care. An 8-year-old spayed female golden retriever was referred to our hospital with a 2-month history of fatigue, weakness, and selective appetite. The dog was diagnosed with chronic lymphocytic leukemia based on blood smear evaluation. Despite treatment, the dog deteriorated and the owners opted for euthanasia. Histopathological examination revealed atypical lymphocytes in the spleen, extramedullary hematopoiesis in the spleen and liver, bone marrow depletion, moderate neutrophilic myocarditis, and acute renal tubular necrosis. In an immunohistochemical panel, cells in the spleen and bone marrow were positive for anti-CD20, anti-CD45, anti-CD3, anti-IBA-1, anti-Ki67, anti-MUM-1, and anti-PAX-5. In conclusion, the leukemia diagnosis was confirmed through reactive histiocytosis, marked extramedullary hematopoiesis, and other clinical and laboratory findings. Herein, we report a case of chronic lymphocytic leukemia in a female dog with clinical, laboratory, and necroscopy findings.

Update on Canine and Human Rabies in a Rabies Endemic Situation in the Republic of Guinea

Each year, rabies kills nearly 60,000 people worldwide. Asia and Africa are the most affected. It is a public health problem because of the number of stray dogs in these nations. In Guinea, rabies is endemic and cases are often under-reported. The objective of this study was to determine the prevalence of rabies among human and animal exposures in Guinea. This was a retrospective analytical study from 2019 to 2021 conducted in the country’s human and veterinary health divisions and, primarily focused on rabies épidémiologique surveillance. The selection was based on human contact with an animal causing behavioural disorders, as well as animals put under observation following aggression on humans. The data were analysed using Epi Info version 21 software. Qualitative variables were expressed as proportions with 95% CIs, quantitative variables as mean ± standard deviation. The chi2 test and the Fischer exact test were used to compare proportions. A total of 85 veterinary facilities were visited and 100 animals registered, of which 93% were reported cases of exposure to canine rabies. One thousand five hundred and thirty-one (1531) people exposed to rabies were recorded between 2019 and 2021. During the period, the prevalence of human rabies was 0.533% [0.042 - 0.064] at 95% CI and that of canine rabies was 0.55% [0.039 - 0.061] at 95% CI. The biting animal was the dog in 97.2% of cases. Subjects over 45 years of age (57.7%) were the most affected. The prevalence of canine rabies was 0.5% compared with 0.533% for human rabies between 2019 and 2021. This low prevalence does not reflect the true extent of the phenomenon in Guinea, as few people consult health structures in the event of dog bites. Regular surveillance of the human and dog population is necessary for better rabies control in Guinea.

Effect of intracoronary autologous bone marrow mononuclear cells transplantation on arrhythmia in canines

Objective To observe the survival and the differentiation of grafted bone marrow cells(BM-MNCs)in host myocardium.To observe whether BM-MNCs transplantation can potentially cause arrhythmia and whether the BM-MNCs transplantation can alter the spatial distribution of connexins,important mediator for arrhythmia gen-

Deworming of stray dogs and wild canines with praziquantel-laced baits delivered by an unmanned aerial vehicle in areas highly endemic for echinococcosis in China

Background:Canines,the definitive hosts for the parasites causing alveolar(AE)and cystic echinococcosis(CE),are the main source of this infections playing the key role in the transmission.The ten-year mortality rate of AE is extremely high(94%)if the patients are not given sustained treatment.The aim of this field study is to explore the possibility of delivery of praziquantel-laced baits using unmanned aerial vehicles(UAVs)aimed at deworming wild canines in the endemic areas.Methods:UAVs were compared to manual bait delivery in the 1-km^(2)test areas followed by testing of canine faeces using an Echinococcus coproantigen ELISA test in the ensuing year.The outcomes of the two approaches were compared with respect to time of delivery and overall cost.Findings:Compared to manual bait delivery,delivery by UAVs saved up to 67%of the overall cost.Three times more staff was needed for the former approach compared to the latter and,time wise,UAV bait delivery saved 350%compared to manual bait delivery on average.With regard to investment needed,the use of UAVs showed an efficiency 2.5 times better than manual bait delivery.Compared to the area served by UAVs,the average positive rate for the canine faecal samples was more than 38%higher in the area served manually.Conclusion:The technique of bait delivery with praziquantel using UAVs for canine deworming has a strong potential with regard to savings of manpower,time and overall cost in areas highly endemic for echinococcosis.

Age-related changes in dispersion of atrial effective refractory period and its ionic mechanism in canines

Atrial fibrillation （AF） is the most common cardiac arrhythmia in man, and epidemiological study has shown the close relationship between the incidence of AF and aging. Although the effects of aging on atrial electrophysiolo have been investigated in humans and animal models, the electrophysiological changes that make the atria of aged individuals more susceptible to AF than those of adults remain poorly understood. AF is a reentrant arrhythmia, and the potential importance of dispersion of effective refractory period （ERP） for the induction and maintenance of reentry has long been recognized. In this study, we measured the dispersion of ERP and the underlying ionic mechanism in the atria of old and adult canines.

Can canines alone be used for age estimation in Chinese individuals when applying the Kvaal method?

Due to the secondary dentin formation,the dental pulp undergoes changes in shape throughout life.Based on this phenomenon,the Kvaal method has been applied to various populations for age estimation,and its usefulness has been verified.When applying the Kvaal method to Chinese subjects,we observed a relatively strong correlation between mandibular canines and age.This study notes the correlation between canines and chronological age and is the first to identify which canine is most closely related to chronological age.In addition,a new,simpler formula is determined based on canines according to Kvaal’s methodology.The radiographs of 360 individuals from northern China were selected,from which the widths and lengths of the pulp from four canines were measured according to the Kvaal method.Next,inter-and intra-observer reliabilities were analyzed in order to assess the repeatability of these measurements.The correlation between measurements and age was examined,and Chinese-specific age estimation formulae were derived.The results revealed that the ratios from the left maxillary canine exhibited the strongest correlation with age compared to the other canines,whereas the left mandibular canine showed the weakest correlation,which may contribute to the overall poor correlation of mandibular canines with age.What’s more,the formula derived from the left maxillary canine in this study displayed the highest coefficients of determination,and the formula derived from all canines showed the lowest residuals.Both of these formulae performed better than the Chinese-specific formula derived from six different types of teeth in our previous study,which had formerly possessed the highest coefficients of determination and the lowest residuals.Thus,we concluded that canines do play an important role in age estimation in the Chinese population,and the correlation between maxillary canines and chronological age is stronger than that of mandibular canines,although no distinct trend as to which side is better correlated with age was established.Going forward,we recommend the analysis of additional samples from different geographical regions and populations to further verify the importance of canines in age estimation.